The Disulfide Bond,but Not Zinc or Dimerization,Controls Initiation and Seeded Growth in Amyotrophic Lateral Sclerosis-linked Cu,Zn Superoxide Dismutase (SOD1) Fibrillation

Aggregation of copper-zinc superoxide dismutase (SOD1) is a defining feature of familial ALS caused by inherited mutations in the sod1 gene, and misfolded and aggregated forms of wild-type SOD1 are found in both sporadic and familial ALS cases. Mature SOD1 owes its exceptional stability to a number of post-translational modifications as follows: formation of the intramolecular disulfide bond, binding of copper and zinc, and dimerization. Loss of stability due to the failure to acquire one or more of these modifications is proposed to lead to aggregation in vivo. Previously, we showed that the presence of apo-, disulfide-reduced SOD1, the most immature form of SOD1, results in initiation of fibrillation of more mature forms that have an intact Cys-57–Cys-146 disulfide bond and are partially metallated. In this study, we examine the ability of each of the above post-translational modifications to modulate fibril initiation and seeded growth. Cobalt or zinc binding, despite conferring great structural stability, neither inhibits the initiation propensity of disulfide-reduced SOD1 nor consistently protects disulfide-oxidized SOD1 from being recruited into growing fibrils across wild-type and a number of ALS mutants. In contrast, reduction of the disulfide bond, known to be necessary for fibril initiation, also allows for faster recruitment during seeded amyloid growth. These results identify separate factors that differently influence seeded growth and initiation and indicate a lack of correlation between the overall thermodynamic stability of partially mature SOD1 states and their ability to initiate fibrillation or be recruited by a growing fibril.     

Manganoporphyrins and ascorbate enhance gemcitabine cytotoxicity in pancreatic cancer

Pharmacological ascorbate (AscH⁻) selectively induces cytotoxicity in pancreatic cancer cells vs normal cells via the generation of extracellular hydrogen peroxide (H₂O₂), producing double-stranded DNA breaks and ultimately cell death. Catalytic manganoporphyrins (MnPs) can enhance ascorbate-induced cytotoxicity by increasing the rate of AscH⁻ oxidation and therefore the rate of generation of H₂O₂. We hypothesized that combining MnPs and AscH⁻ with the chemotherapeutic agent gemcitabine would further enhance pancreatic cancer cell cytotoxicity without increasing toxicity in normal pancreatic cells or other organs. Redox-active MnPs were combined with AscH⁻ and administered with or without gemcitabine to human pancreatic cancer cell lines, as well as immortalized normal pancreatic ductal epithelial cells. The MnPs MnT2EPyP (Mn(III)meso-tetrakis(N-ethylpyridinium-2-yl) porphyrin pentachloride) and MnT4MPyP (Mn(III)tetrakis(N-methylpyridinium-4-yl) porphyrin pentachloride) were investigated. Clonogenic survival was significantly decreased in all pancreatic cancer cell lines studied when treated with MnP + AscH⁻ + gemcitabine, whereas nontumorigenic cells were resistant. The concentration of ascorbate radical (Asc^•−, an indicator of oxidative flux) was significantly increased in treatment groups containing MnP and AscH⁻. Furthermore, MnP + AscH⁻ increased double-stranded DNA breaks in gemcitabine-treated cells. These results were abrogated by extracellular catalase, further supporting the role of the flux of H₂O₂. In vivo growth was inhibited and survival increased in mice treated with MnT2EPyP, AscH⁻, and gemcitabine without a concomitant increase in systemic oxidative stress. These data suggest a promising role for the use of MnPs in combination with pharmacologic AscH⁻ and chemotherapeutics in pancreatic cancer.     

禾谷镰刀菌粗毒素对不同小麦品种幼苗MDA含量和SOD、PAL活性的影响

用禾谷镰刀菌粗毒素对赤霉病抗性不同的6个小麦品种(系)幼苗进行处理,在1h~10d内测定各品种(系)的MDA含量、SOD及PAL活性增率的变化.结果显示:禾谷镰刀菌粗毒素处理后,抗病和中抗品种(系)幼苗的MDA含量增率低于感病品种(系),且增加速度慢;抗病和中抗品种(系)幼苗的SOD活性在胁迫1h后迅速升高,其活性增率高于感病品种(系);抗、感品种(系)PAL活性均升高,但抗病品种(系)PAL活性增加快,且活性增率高于感病品种(系).研究表明,小麦幼苗的SOD、PAL活性增率与品种抗病性呈正相关,MDA含量增率与品种抗病性呈负相关.     

菠菜铁型超氧化物歧化酶的纯化及性质

用聚丙烯胺梯度凝胶电泳法检测出菠菜ＳＯＤ同工酶谱带中含３条Ｆｅ－ＳＯＤ活性带,菠菜叶Ｆｅ－ＳＯＤ粗提取液经硫酸铵分部沉淀,ＤＥＡＥ－纤维素－Ａ５２和ＳｅｐｈａｄｅｘＧ－１００柱层析,纯化出单一的Ｆｅ－ＳＯＤ活性带,纯化酶的分子量为４２．６ｋＤ,亚基分子量为２１ｋＤ。对金属元素的分析表明,该酶每分子含２．６个Ｆｅ原子,该酶紫外区最大吸收峰为２７８ｎｍ,等电点为４．６,氨基酸组成和其它来源的Ｆｅ－ＳＯ     

不同水温游泳对小鼠行为,记忆和脂质过氧化影响差异的研究

目的 :探讨不同强度应激对小鼠衰老改变影响的差异。方法 :用D 半乳糖进行衰老诱导的小鼠同步进行不同水温 (12℃、18℃、2 5℃ )游泳应激训练 ,连续六周后 ,进行开场行为、学习记忆能力、半脑MDA含量及海马SOD活性测定。结果 :D 半乳糖诱导组 (D)较对照组 (C)探究行为减少 ,获得性行为能力减退 (P <0 .0 1) ,且脑内脂褐素含量明显增加 (P <0 .0 5 ) ;18℃及 2 5℃水温应激训练有对抗D 半乳糖所致行为和学习记忆能力减退的作用 (P <0 .0 1) ,同时脑中抗氧化能力提高 (P <0 .0 5 ) ,而 12℃水温组学习记忆能力进一步减退 ,海马SOD活性进一步下降(P <0 .0 5 ) ,且各项行为能力及SOD活性与 18℃和 2 5℃组均有显著差异 (P <0 .0 1)。结论 :不同水温游泳应激对机体的影响具有双重性 ,适量的应激刺激可逆转机体的衰老改变 ,提高动物的学习记忆能力 ,而过强的应激刺激有加速衰老进程的倾向。两种应激对机体抗氧化能力的不同改变可能是引起这一差异的原因。     

春玉米叶片衰老中激素变化,Ca^2＋跨膜运输和膜脂过氧化三者之间的关系

通过离体玉米（ZeamaysL．）叶片培养和叶肉质膜微囊45Ca2 吸收等实验,探索春玉米叶片衰老过程中激素变化、Ca2 跨膜运输及膜脂过氧化三者之间的联系。结果认为,玉米叶片衰老的可能过程首先是内源激素含量变化,继而影响到Ca2 跨膜运输,进而导致膜脂过氧化,由此引起叶绿素和蛋白质降解。     

转Bt基因水稻对稻纵卷叶螟幼虫体内三种保护酶活性的影响

用转Cry1Ab/Cry1Ac基因水稻的叶片饲喂稻纵卷叶螟Cnaphalocrocis medinalis (Guenée)幼虫,采用酶活力测定方法研究了转基因水稻对稻纵卷叶螟幼虫体内3种保护酶（SOD,CAT和POD）活性的影响。结果表明：取食转基因水稻叶片4 h后,幼虫体内SOD酶活性比对照提高了91.5%,与对照有显著差异;36 h后活性达到最大值,但与对照差异不显著。取食转基因水稻叶片24 h后,幼虫体内CAT酶活性达到最大值且高于对照,但与对照差异不显著;48 h后酶活性受到显著抑制,与对照差异显著。取食转基因水稻叶片12 h后,幼虫体内POD酶活性达到最大值,与对照差异不显著;48 h后酶活性逐渐下降达到最小值,比对照下降68.05%。实验同时测定了幼虫体内及其粪便中Bt毒蛋白含量的变化,结果表明取食转基因水稻叶片12 h后,随着大量取食进入体内的毒蛋白随粪便排出,幼虫体内的毒蛋白含量一直低于粪便中的含量,且在24 h时两者差异达到最大。由于Bt毒蛋白在幼虫体内的积累,扰乱了稻纵卷叶螟幼虫体内SOD,CAT和POD 3种保护酶的动态平衡,使虫体内自由基的清除遇到障碍,从而对其产生毒害作用。     

中华通草蛉自然越冬成虫在长、 短光周期下滞育解除中体内相关酶活力变化

光周期信号在昆虫的环境适应中发挥着重要作用, 昆虫能够通过感受光周期的变化来调节体内生理生化过程, 以适应环境的变化。为明确光周期对中华通草蛉Chrysoperla sinica越冬成虫滞育解除过程中酶活力的影响, 本研究测定了长光周期（15L∶9D）和短光周期（9L∶15D）条件下, 成虫体内过氧化氢酶（CAT）、 超氧化物歧化酶（SOD）、 Na⁺K⁺-ATP酶和乳酸脱氢酶（LDH） 4种重要酶活力的变化。结果表明： 中华通草蛉雌、 雄成虫CAT活性在长光周期处理5 d达最高值后呈下降趋势; 短光周期处理CAT活性在处理5 d达最高值, 且高于长光周期处理, 在处理10 d迅速下降至最低值, 且均显著低于长光周期处理的CAT活性（P=0.005）, 后迅速上升并在处理15 d （P<0.05）和20 d （P<0.005）活性显著高于长光周期处理。雌、 雄成虫SOD活性在长光周期下处理10 d达最高值, 且显著高于对照（P<0.001）, 且除处理5 d雄虫SOD活性与对照无显著性差异外（P=0.558）其余处理活性均显著低于对照（P<0.05）。雌成虫长光周期处理5 d 的SOD活性显著低于短光周期（P<0.001）, 其余处理活性均显著高于短光周期; 雄成虫长光周期下处理的SOD活性均高于短光周期下, 且处理5 d （P=0.04）, 15 d （P<0.001）和20 d （P=0.003）的活性差异显著。两种光周期条件下雌、 雄成虫Na⁺K⁺-ATP酶活性随处理时间延长呈上升-下降-上升趋势, 且均显著高于处理0 d成虫酶活性（P<0.001）; 短光周期处理不同时间Na⁺K⁺-ATP酶活性均高于长光周期处理, 且除雄成虫处理15 d无显著性差异（P=0.142）外, 其余均差异显著（P<0.05）。两种光周期条件下雌、 雄成虫LDH活性随处理时间延长呈下降趋势, 且均显著低于对照（P<0.001）。中华通草蛉越冬成虫在长、 短两种光周期条件下体内酶活力的差异可能是影响两种光周期下成虫滞育解除过程中体内不同生化物质含量与生殖状态的重要因子。